Bacteriolytic complex, method for producing said complex and strain for carrying out said method

Abstract

The inventions relate to the field of medicine, veterinary and biotechnology and may be used for production of a drug for medical and veterinary purposes. A bacteriolytic complex, produced by the bacterium Lysobacter sp. XL1, has been proposed, which contains bacteriolytic enzymes (muramidase, muramoylalanineamidase, endopeptidase, a bacteriolytic enzyme with a molecular weight of about 22 kDa), protease, polysaccharide, and ballast components. The method of production of the bacteriolytic complex includes cultivation of the strain-producer on a nutrient medium containing glucose, peptone, yeast extract or yeast autolysate, phosphate salts of sodium and potassium, magnesium sulfate, potassium chloride, iron sulfate, and water.

Description

TECHNICAL FIELD [0001]The inventions relate to the field of biotechnology, medicine, and veterinary, more precisely concerning a bacteriolytic complex, and may be used for production of a drug for medicine and veterinary.BACKGROUND[0002]It is known that bacteriolytic preparations are used as drugs in treatment for a number of diseases caused by pathogenic microflora, including that resistant to antibiotics.[0003]The preparation having bacteriolytic effect—lysozyme is known (Egorov N. S. Bases of the doctrine of antibiotics, 1979, “Vysshaya shkola”, p. 347–349). Lysozyme of egg protein is active against a narrow spectrum of microorganisms, limited mainly by micrococci. By specificity of action on microbial cell wall peptidoglycan, lysozyme is a muramidase, i.e., lyzes the bond between muramic acid and glucosamine with the formation of fragments with muramic acid on the reducing end.[0004]That is why lysozyme lyzes the narrow spectrum of gram-positive microorganisms. In particular, it...

AI Technical Summary

Benefits of technology

[0048]The technical result expected to obtain if the invention is realized, is an increase of the quantity and the content of bacteriolytic enzymes in culture liquid of the bacterium-producer, an increase of product yield, optimization of the composition of end product (reduction of the content of inactive proteins in the end product), acceleration of production process of finished product.

[0049]In this context, the price of the product drops and product production process accelerates.

[0060]With the proposed components of the nutrient medium and their proportion, the price of the medium is lower and the content of bacteriolytic enzymes in culture liquid of the producer is raised from 30–60 LE / ml (with the known methods) to 70–90 LE / ml, and the content of inactive proteins in culture medium decreases (FIG. 1).

[0071]A change of the procedure of precipitation, namely the exclusion of fractional precipitation by acetone (ru patent 1549227) or by alcohol (ru patent 1755581) from the procedure of purification, reduces duration of the process of the end product obtaining and increases the yield of the target product from 20–50% (with the known methods) to 80–100% of the content in culture liquid.

Problems solved by technology

Besides, like most enzyme preparations, lyzozyme is not stable enough.

However, it is unstable and applied in laboratory practice only.

The disadvantage of this complex is that its bacteriolytic activity is not high enough due to the presence of ballast components and because nearly the half of the bacteriolytic enzymes are denatured.

Besides, the ballast components have no medicinal effect but may cause allergic reactions.

Both the content of bacteriolytic enzymes in culture liquid and the yield of finished preparation are not high enough at realization of this method.

The yield of end product is not high enough—about 20% of the lytic complex content in culture liquid.

The above method has a disadvantage that the protein-vitamin concentrate is currently not produced by domestic industry, because its production is ecologically unsafe; the use of expensive imported bactopeptone Difco (USA) as a medium component is unpractical, because it significantly increases the cost of the final product.

Besides, the use of ethanol (RU 1755581) and acetone (RU 1549227) for precipitation of the enzymes from culture liquid of the producer results in irreversible denaturing of about the half of bacteriolytic enzymes determining the therapeutic effect of the drug, which are present in the culture liquid (according to the results of bacteriolytic activity measurement and electrophoretic analysis) and, consequently, in a considerable decrease in the product yield (FIG. 1).

Finished lysoamidase preparations produced by the known methods contain a significant amount of minor components—ballast proteins (FIG. 1), which have no medicinal effect but may cause an allergic reaction.

The disadvantage of this strain-producer is that, due to the presence of minor components in the produced complex, bacteriolytic activity is insufficiently high.

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